HCT-8 [HRT-18] (ATCC® CCL-244)

Organism: Homo sapiens, human  /  Tissue: colon  /  Disease: ileocecal colorectal adenocarcinoma

Organism Homo sapiens, human
Tissue colon
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease ileocecal colorectal adenocarcinoma
Age 67 years adult
Gender male
Storage Conditions liquid nitrogen vapor phase
Clinical Data
male
Genes Expressed
carcinoembryonic antigen (CEA) 0.5 ng/106 cells/10 days; alkaline phosphatase. The cells are positive for keratin by immunoperoxidase staining.
Cellular Products
carcinoembryonic antigen (CEA) 0.5 ng/10 exp6 cells/10 days; alkaline phosphatase; keratin
Tumorigenic Yes
Effects

Yes, in nude mice

Tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 107 cells

 

Comments
The HCT-8 line is identical to the HRT-18 cell line.

The cells are positive for keratin by immunoperoxidase staining.

 

Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: horse serum to a final concentration of 10%.
Subculturing

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which  contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.  Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. 
  5. To remove trypsin-EDTA solution, transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to10 minutes.
  6. Discard supernatant and resuspend cells in fresh growth medium.
  7. Add appropriate aliquots of the cell suspension to new culture vessels.
  8. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended

Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X,Y
CSF1PO: 12
D13S317: 8,11
D16S539: 12,13
D5S818: 13
D7S820: 10,12,11.3
THO1: 7,9.3
TPOX: 8,11
vWA: 18,19
Isoenzymes
AK-1, 1
ES-D, 1-2
G6PD, B
GLO-I, 2
Me-2, 1
PGM1, 1
PGM3, 1
Name of Depositor A Sobrero
References

Tompkins WA, et al. Cultural and antigenic properties of newly established cell strains derived from adenocarcinomas of the human colon and rectum. J. Natl. Cancer Inst. 52: 1101-1110, 1974. PubMed: 4826581

Nelson-Rees WA, et al. Distinctive banded marker chromosomes of human tumor cell lines. Int. J. Cancer 16: 74-82, 1975. PubMed: 1058173

White LJ, et al. Attachment and entry of recombinant norwalk virus capsids to cultured human and animal cell lines. J. Virol. 70: 6589-6597, 1996. PubMed: 8794293

Basic Documentation
References

Tompkins WA, et al. Cultural and antigenic properties of newly established cell strains derived from adenocarcinomas of the human colon and rectum. J. Natl. Cancer Inst. 52: 1101-1110, 1974. PubMed: 4826581

Nelson-Rees WA, et al. Distinctive banded marker chromosomes of human tumor cell lines. Int. J. Cancer 16: 74-82, 1975. PubMed: 1058173

White LJ, et al. Attachment and entry of recombinant norwalk virus capsids to cultured human and animal cell lines. J. Virol. 70: 6589-6597, 1996. PubMed: 8794293